DETAILED ACTION
Notice of Pre-AIA or AIA Status
The present application, filed on or after March 16, 2013, is being examined under the first inventor to file provisions of the AIA .
Status of the Claims
Claims 1-27 are currently pending.
Claims 20-22 and 25-27 have been withdrawn from further consideration pursuant to 37 CFR 1.142(b) as being drawn to a nonelected Invention, there being no allowable generic or linking claim.
Claims 1-19 and 23-24 have been considered on the merits.
Maintained Rejections Necessitated by Amendment
Claim Rejections - 35 USC § 103
In the event the determination of the status of the application as subject to AIA 35 U.S.C. 102 and 103 (or as subject to pre-AIA 35 U.S.C. 102 and 103) is incorrect, any correction of the statutory basis (i.e., changing from AIA to pre-AIA ) for the rejection will not be considered a new ground of rejection if the prior art relied upon, and the rationale supporting the rejection, would be the same under either status.
The following is a quotation of 35 U.S.C. 103 which forms the basis for all obviousness rejections set forth in this Office action:
A patent for a claimed invention may not be obtained, notwithstanding that the claimed invention is not identically disclosed as set forth in section 102, if the differences between the claimed invention and the prior art are such that the claimed invention as a whole would have been obvious before the effective filing date of the claimed invention to a person having ordinary skill in the art to which the claimed invention pertains. Patentability shall not be negated by the manner in which the invention was made.
This application currently names joint inventors. In considering patentability of the claims the examiner presumes that the subject matter of the various claims was commonly owned as of the effective filing date of the claimed invention(s) absent any evidence to the contrary. Applicant is advised of the obligation under 37 CFR 1.56 to point out the inventor and effective filing dates of each claim that was not commonly owned as of the effective filing date of the later invention in order for the examiner to consider the applicability of 35 U.S.C. 102(b)(2)(C) for any potential 35 U.S.C. 102(a)(2) prior art against the later invention.
Claims 1-7, 10-12, 15-16, and 19 are rejected under 35 U.S.C. 103 as being unpatentable over Campbell et al (US20030175410A1) in view of Birey et al (Nature, 2017).
With regards to claim 1, Campbell teaches a method of generating tissue by depositing through bioprinting tissue precursor cells ([0005]/[0008]/claim 64). Further, Campbell teaches that the bioink is deposited a plurality of times ([0155]). The bioink is taught to contain hydrogel ([0009]), hyaluronic acid ([0169]), fibrinogen ([0088]), thrombin ([0088]), and a protease inhibitor ([0109]) as required by claims 2, 3, and 4. Further, the hydrogel is crosslinked after each depositing layer as required by claim 5 ([0010]/[0206]). The crosslinking agent is taught to be CaCl2, thrombin and transglutaminase as required by claim 6 ([0010]/[0220]). The plurality of bands (layers) are taught to be printed/deposited horizontally next to each other on the suitable surface as required by claim 7 ([0155]).
Campbell does not teach that the precursor cells in the bioink are glutamatergic cortical progenitor cells and GABAergic interneuron progenitor cells and maturing the cells to form a neural tissue construct as required by claim 1. Campbell does not teach that a functional neural network is formed in about 2-4 weeks as required by claims 15 and 16. Campbell does not teach that the Glutamatergic cortical neural progenitors and GABAergic interneuron progenitors are 21 day hPSC derived cells as required by claims 10 and 11. Campbell does not teach the inclusion of astrocyte progenitor cells as required by claim 12. Campbell does not teach the maturation in basal medium with supplements as required by claim 19.
However, Birey teaches a method of generating a neural tissue construct where a 3-dimensional (3D) spheroid culture of human pluripotent stem cells (hPSC) derived neural cells are differentiated into glutamatergic cortical interneurons (pg. 56, column 1, para. 1) and GABAergic neurons (pg. 55, column 2, para 1). Further, Birey teaches that in their spheroid model, interneurons functionally integrate with glutamatergic neurons to form a microphysiological system (abstract). Birey also teaches that the spheroid migrates to form a functional neural network in 2 weeks as required by claims 15 and 16 (pg. 55, column 2, para. 1). The cells taught by Birey are in vitro differentiated and tested at day 25 and further cultured for 2 weeks, which meets the limitations of using 21-day differentiated cells as required by claims 10 and 11 (pg. 54, column 2, para. 1). Birey teaches the spontaneous presence of astrocytes in the spheroid cultures, which meets the limitation of containing astrocyte progenitors as required by claim 12 (pg. 55, column 1, para. 2). The spheroid culture is taught to be performed in a neuronal base medium containing B-27 supplement, brain derived neurotrophic factor (BDNF), and a ROCK inhibitor as required by claim 19 (Methods, pg. 60, column 1, para. 3). In further support, Campbell teaches the method where the bio-inks contain cells from the brain (0127).
One of ordinary skill in the art would find it obvious at the time of the effective filling date to combine the bioink printing method taught by Campbell with the neural cell types taught by Birey to arrive at the instant invention. One of ordinary skill in the art would be motivated to make this modification because Birey teaches that in their spheroid model, interneurons functionally integrate with glutamatergic neurons to form a microphysiological system (abstract). One of ordinary skill in the art would have a reasonable expectation of success when combining Campbell with Birey because Campbell teaches that the method can be performed with tissue precursor cells and Birey teaches that the 3D culture of neural precursor cells results in a functional neural model.
Therefore, the invention as a whole was prima facie obvious to one of ordinary skill in the art at the effective time the invention was filed, especially in the absence of evidence to the contrary.
Claims 8-9, 13, and 17-18 are rejected under 35 U.S.C. 103 as being unpatentable over Campbell et al (US20030175410A1) and Birey et al (Nature, 2017) as applied to claims 1-7, 10-12, 15-16, and 19 above, and further in view of Murphy et al (US20130190210A1).
With regards to claim 8, Campbell and Birey teach the limitations of the independent claim 1.
Campbell and Birey do not teach that the thickness or size of the printed bands is about 50 μm thick as required by claim 8.
However, Murphy teaches a method of bioprinting layers of cells in which the band or layer of cells that are printed can be anywhere from 3-500 μm thick ([0073]).
One of ordinary skill in the art would find it obvious at the time of the effective filling date to combine method using bioink taught by Campbell and Birey with the band thickness taught by Murphy to arrive at the instant invention. One of ordinary skill in the art would be motivated to make this combination because bands of 50 μm thick were known to be bioprinted as taught by Murphy. Additionally, one of ordinary skill in the art would recognize that the thickness of the bands in a bioprinted tissue is a result effective variable and that the thickness of the bands in a bioprinted tissue would be matter of routine optimization depending on the application and desired tissue. One of ordinary skill in the art would have a reasonable expectation of success because Murphy teaches a wide range of thicknesses which are applicable for bioprinting.
Birey teaches the formation of a functional neural network in 2 weeks as required by claims 17 and 18 (pg. 55, column 2, para. 1). Campbell and Birey do not teach that the ratio of glutamatergic cortical progenitor cells and the GABAergic interneuron progenitor cells are present in the mixture at a ratio of 4:1 as required by claim 9. Campbell and Birey does not teach that the ratio of glutamatergic cortical progenitor cells, GABAergic interneuron progenitor cells, and astrocyte progenitor cells are present in the mixture at a ratio of 5:1:4 as required by claim 13.
However, Murphy teaches the use of neural cells in the bioink ([0004]). Further, Murphy teaches that the cell types and sources of cells used to fabricate one or more tissues are to be selected based on a specific research goal or objective and that the types and ratios of cells depend on the desired tissue formation as required by claims 9 and 13 ([0128]).
One of ordinary skill in the art would find it obvious at the time of the effective filling date to combine method using bioink taught by Campbell and Birey with the adjustable cell ratios taught by Murphy to arrive at the instant invention. One of ordinary skill in the art would be motivated to make this combination Murphy teaches that the cell types and sources of cells used to fabricate one or more tissues are to be selected based on a specific research goal or objective and that the types and ratios of cells depend on the desired tissue formation ([0128]). One of ordinary skill in the art would have a reasonable expectation of success because Murphy teaches selecting cell types and ratios based on the research goal or objective for bioprinting.
Therefore, the invention as a whole was prima facie obvious to one of ordinary skill in the art at the effective time the invention was filed, especially in the absence of evidence to the contrary.
Claim 14 is rejected under 35 U.S.C. 103 as being unpatentable over Campbell et al (US20030175410A1) and Birey et al (Nature, 2017) as applied to claims 1-7, 10-12, 15-16, and 19 above, and further in view of Colonna et al (Annu Rev Immunol, 2017).
With regards to claim 14, Campbell and Birey teach the limitations of the independent claims 1 and 12.
Campbell and Birey do not teach that the mixture comprises microglial cells.
However, Colonna teaches that microglial cells are resident cells of the brain that regulate brain development, maintenance of neural networks, and injury repair (abstract).
One of ordinary skill in the art would find it obvious at the time of the effective filling date to combine method using bioink taught by Campbell and Birey with the microglial cells taught by Colonna to arrive at the instant invention. One of ordinary skill in the art would be motivated to make this combination, because Colonna teaches that microglial cells are resident cells of the brain that regulate brain development, maintenance of neural networks, and injury repair (abstract) and it would be obvious to include cells found in nature in the desired tissue type, brain tissue. One of ordinary skill in the art would have a reasonable expectation of success because including microglial cells will better imitate the natural environment of the brain.
Therefore, the invention as a whole was prima facie obvious to one of ordinary skill in the art at the effective time the invention was filed, especially in the absence of evidence to the contrary.
Claims 23 and 24 are rejected under 35 U.S.C. 103 as being unpatentable over Campbell et al (US20030175410A1) and Birey et al (Nature, 2017) as applied to claims 1-7, 10-12, 15-16, and 19 above, and further in view of Yger et al (Front. Behavioral Neuroscience, 2011), Kitagawa et al (Journal of Neurosci., 2017) and Deo et al (Neurobiology, 2018).
With regards to claim 23, Campbell and Birey teach the limitations of the independent claim 1.
Campbell and Birey do not teach that the mixture comprises DARPP32+ striatal medium spiny neurons as required by claim 23.
However, Yger teaches that DARPP32+ is highly enriched in striatal medium spiny neurons. Further, Yger teaches that the major function of DARPP-32 is to increase the reliability of signal processing in striatal medium-size spiny neurons (pg. 4, column 2, para 1).
One of ordinary skill in the art would find it obvious at the time of the effective filling date to combine method using bioink taught by Campbell and Birey with the Striatal medium spiny neurons taught by Yger to arrive at the instant invention. One of ordinary skill in the art would be motivated to make this combination Yger teaches that the major function of DARPP-32 is to increase the reliability of signal processing in striatal medium-size spiny neurons (pg. 4, column 2, para 1) and it would be obvious to include cells found in nature in the desired tissue type, brain tissue. One of ordinary skill in the art would have a reasonable expectation of success because including DARPP-32+ striatal medium spiny neurons will better imitate the natural environment of the brain.
Birey teaches the use of a cell marker DLX1-eGFP to mark GABAergic cells for characterization (Fig 1j, pg. 55, column 1, para 1). Campbell and Birey do not teach that the mixture comprises cortical neurons as required by claim 23. Campbell, Birey, and Yger do not teach that the cortical neurons in the neural tissue express ChR2-EYFP nor that the striatal neurons express a red calcium indicator jRGECO1b as required by claim 24.
However, Kitagawa teaches monitoring the CREB behavior of cortical neurons using ChR2-EYFP-expressing cortical neurons as required by claim 23 and 24 (Fig 5A).
One of ordinary skill in the art would find it obvious at the time of the effective filling date to combine method using bioink taught by Campbell and Birey with ChR2-EYFP-expressing cortical neurons taught by Kitagawa to arrive at the instant invention. One of ordinary skill in the art would be motivated to make this combination because it is obvious to include cells with detectible markers so that the different cell types can be distinguished as done by Birey through the use of a cell marker DLX1-eGFP to mark GABAergic cells for characterization (Fig 1j, pg. 55, column 1, para 1). One of ordinary skill in the art would have a reasonable expectation of success because Kitagawa teaches the successful monitoring the CREB behavior of cortical neurons using ChR2-EYFP-expressing cortical neurons (Fig 5A).
Campbell, Birey, Yger, and Kitagawa do not teach that the striatal neurons express a red calcium indicator jRGECO1b.
However, Deo teaches a red calcium indicator, jRGECO1b, as a small molecule probe (pg. 105, column 2, para 2). Further Deo teaches that more sophisticated sensing schemes can be accessed by developing molecular logic gate systems with small molecule probes and that this allows time-gated marking of active neurons (pg. 105, column 2, para 2).
One of ordinary skill in the art would find it obvious at the time of the effective filling date to combine method using bioink taught by Campbell, Birey, Yger and Kitagawa with the jRGECO1b small molecule probe to arrive at the instant invention. One of ordinary skill in the art would be motivated to make this combination because it is obvious to include cells with detectible markers so that the different cell types can be distinguished as done by Birey through the use of a cell marker DLX1-eGFP to mark GABAergic cells for characterization (Fig 1j, pg. 55, column 1, para 1) and because Deo teaches that more sophisticated sensing schemes can be accessed by developing molecular logic gate systems with small molecule probes and that this allows time-gated marking of active neurons (pg. 105, column 2, para 2). One of ordinary skill in the art would have a reasonable expectation of success because Birey, Kitagawa, and Deo teach the successful labeling of cells using various markers.
Therefore, the invention as a whole was prima facie obvious to one of ordinary skill in the art at the effective time the invention was filed, especially in the absence of evidence to the contrary.
Response to Arguments
Applicant's arguments filed 11/20/2025 have been fully considered but they are not persuasive.
Applicant argues (Remarks, pg. 6, last para) that a person of ordinary skill in the art would not have been motivated to combine Campbell and Birey to arrive at the claimed invention because 1) Campbell’s teachings do not aid a skilled artisan in the application of a bioink printing technology to specialized and complex neural progenitor cells and 2) Birey’s teachings are fundamentally different from the claimed invention because the methodology employs spheroids which are not individual neural progenitor cells. Applicant claims that the disclosure of Campbell alone does not teach the requirements of the specific invention and states “[t]he closest Campbell gets to teaching the use of neural tissue progenitors is an unsupported statement that the model could include cells from the brain to serve as treatment for various nervous system disorders and diseases” (Remarks, pg. 7, para 1). Applicant also states that Birey does not cure the alleged deficiencies of Campbell because “Birey does not teach a 3D culture of individual neural progenitor cells migrating to form a neural tissue; rather, it teaches the formation of specific neural tissue constructs from specifies neural tissue cell aggregates” (Remarks, pg. 7, para 2).
In response, it appears that applicant’s arguments attacks the references individually by stating that the method of generating biomimetic scaffolds using bioink printing methods Campbell does not teach the exact progenitor cells and that the progenitor cells of Birey are not cultured in the bioink method of culture. Campbell teaches a method of generating biomimetic scaffolds using bioink printing methods and explicitly teaches that the methods can be used for treatment of the central nervous system including striatal brain tissue for Huntington’s disease and dopamine-rich brain tissue and cells for Parkinson’s disease ([0164]). The striatum is known for being a dopamine rich brain region composed primarily of GABAergic medium spiny neurons, therefore Campbell is at the very least supporting the use of the bioink printing method employing neural cells generally.
As stated in the previously presented rejection, “Birey teaches a method of generating a neural tissue construct where a 3-dimensional (3D) spheroid culture of human pluripotent stem cells (hPSC) derived neural cells are differentiated into glutamatergic cortical interneurons (pg. 56, column 1, para. 1) and GABAergic neurons (pg. 55, column 2, para 1)”. Claim 1 requires that the mixture being bioprinted comprises glutamatergic cortical progenitor cells and GABAergic interneuron progenitor cells. Therefore, the “progenitor cell” as claimed are met by the human pluripotent stem cell derived neural cells which are the progenitor cell of both the glutamatergic cortical interneurons and GABAergic neurons. Applicant appears to be arguing that the resulting spheroids of Birey are not “individual progenitor cells” however, the resulting spheroids are not being relied upon to meet the limitation of the progenitor cells, rather the starting material of the human pluripotent stem cells (hPSC) derived neural cells of Birey is being relied upon. Additionally, due to the combination of Campbell and Birey, Birey is not being relied upon to teach the method of culture rather only to teach the starting material and capabilities of the cells when grown in 3D culture. The 3D culture of Birey may not be a bioink-utilizing 3D method, however Campbell is relied upon to teach this method. One of ordinary skill in the art would be motivated to combine Campbell and Birey because Campbell teaches the method can be used for neural cell related cultures and Birey a starting material which meets the limitations of the claimed glutamatergic cortical progenitor cells and GABAergic interneuron progenitor cells, which results in interneurons functionally integrating with glutamatergic neurons to form a microphysiological system (abstract).
Therefore, in response to applicant's arguments against the references individually, one cannot show nonobviousness by attacking references individually where the rejections are based on combinations of references. See In re Keller, 642 F.2d 413, 208 USPQ 871 (CCPA 1981); In re Merck & Co., 800 F.2d 1091, 231 USPQ 375 (Fed. Cir. 1986). Therefore, the arguments are not found persuasive.
Applicant argues (Remarks, pg. 7, last para spanning pg. 8) that Birey teaches away from the combination of Campbell and Birey because the discussion focuses on observation of saltatory migration. Applicant states “[t]his distinction is critical as Birey studies how differentiated cell types and tissue behave within a biologically inspired context, while the present invention actively engineers that context through a controlled fabrication process to build a functional tissue construct from its foundational cellular components” (Remarks, pg. 8, para 1). Additionally, Applicant states “Birey does not provide any further suggestion that neural progenitors would be obvious to use in Campbell’s model either” (pg. 8, para 2).
In response to Applicant’s argument that Birey teaches away from the combination, it is not clear how Birey supports a conclusion of teaching away. Birey employs a starting material which meets the claim limitations of the progenitor cells as described in detail at point 13 above and the observational data that Birey collects does not appear to teach away from the use of the starting cell materials, which is what Birey is mainly relied upon to demonstrate. Applicant states that Birey studies how the cells behave in a biologically inspired context and that the present invention actively engineers “that context”, which is meant to support a conclusion of Birey teaching away. It is not clear how Birey can teach away from a biologically inspired context that the instant invention is actively engineering. It appears applicant is considering each reference individually and is not considering the combination of Campbell and Birey, which would result in the same “actively engineered” biological context that is claimed. Therefore, the arguments are not found persuasive.
Applicant argues (Remarks, pg. 8 last para spanning pg. 9, para 2) that the combination of Campbell, Birey, and Murphy would not be obvious because Murphy discloses broad cell types and does not remedy the alleged deficiencies of the combination of Campbell and Birey.
In response, this argument is not found persuasive because the alleged deficiencies of Campbell and Birey are addressed above and Murphy is not relied upon to teach exact cell types. Rather Murphy was relied upon to teach that it would be obvious to optimize the ratio between cell types based on the desired tissue formation. Therefore, the arguments are not found persuasive.
Applicant argues (Remarks, pg. 9-10) that the cited references Colonna, Yger, Kitagawa and Deo do not cure the alleged deficiencies of Campbell and Birey.
In response, these alleged deficiencies have been addressed above at point 13 and therefore the argument is not found persuasive.
Conclusion
No claims are allowed.
THIS ACTION IS MADE FINAL. Applicant is reminded of the extension of time policy as set forth in 37 CFR 1.136(a).
A shortened statutory period for reply to this final action is set to expire THREE MONTHS from the mailing date of this action. In the event a first reply is filed within TWO MONTHS of the mailing date of this final action and the advisory action is not mailed until after the end of the THREE-MONTH shortened statutory period, then the shortened statutory period will expire on the date the advisory action is mailed, and any nonprovisional extension fee (37 CFR 1.17(a)) pursuant to 37 CFR 1.136(a) will be calculated from the mailing date of the advisory action. In no event, however, will the statutory period for reply expire later than SIX MONTHS from the mailing date of this final action.
Examiner Contact Information
Any inquiry concerning this communication or earlier communications from the examiner should be directed to CONSTANTINA E STAVROU whose telephone number is (571)272-9899. The examiner can normally be reached M-F 8:00-5:00.
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CONSTANTINA E. STAVROU
Examiner
Art Unit 1632
/ANOOP K SINGH/Primary Examiner, Art Unit 1632